Helped by benefactor Anna Laura Kidder who saw his potential, Hairston graduated from Tufts University, near Boston, Mass. in 1929. He was one of the first black students admitted to Tufts.

He sang with the Hall Johnson Choir in Harlem for a time but was nearly fired from the all black choir because he had difficulty with the rural dialects that were used in some of the songs. He had to shed his Boston accent and relearn the country speech of his parents and grandparents. (Johnson had reportedly told him, "We're singing ain't and cain't and you're singing shahn't and cahn't and they don't mix in a spiritual."[3]) The Hall Johnson Choir performed in many Broadway shows including Green Pastures. In 1936, they were asked to go to Hollywood to sing for the film,Green Pastures. At that time, a Russian composer, Dimitri Tiomkin, heard Jester and invited him to collaborate with him. This led to a thirty-year collaboration during which time Jester arranged and collected music for the movies. He also wrote and arranged spirituals for Hollywood films as well as for high school and college choirs around the country.[citation needed]

In 1961, the US State Department appointed Jester Hairston as Goodwill Ambassador. He traveled all over the world teaching and performing the folk music of the slaves. No matter where Jester performed in the US, he checked the phone book for Hairstons and was responsible for reuniting people on his family tree, both black and white. He composed more than 300 spirituals. All of his research and work has been documented for history. He was the recipient of four honorary doctorates, including an honorary doctorate from The University of Massachusetts in 1972, and another in music from Tufts in 1977.[4][5]

In his later years, Hairston served as a cultural ambassador for American music, traveling to numerous countries with choral groups that he had assembled. In 1985 he took the Jester Hairston Chorale, a multi-racial group, to sing in the People's Republic of China, at a time when foreign visitors were still quite rare in that country.

Hairston died in Los Angeles of natural causes in 2000 at age 98. Born in 1901, Hairston's life spanned each year of the 20th century. For his contribution to the television industry, Hairston has a star on the Hollywood Walk of Fame located at 6201 Hollywood Blvd. He is interred at Inglewood Park Cemetery, Inglewood, California.